Or indeed support surreal structures :-))
(PMID: 11853672 vs PMID: 14749821 and so on)
------------------------------------------
A. Radu Aricescu, PhD
University Research Lecturer
MRC Career Development Award Fellow
University of Oxford
Wellcome Trust Centre for Human Genetics
Division of Structural Biology
Roosevelt Drive, Oxford OX3 7BN
United Kingdom
Phone: +44-1865-287564
Fax: +44-1865-287547
---- Original message ----
>Date: Wed, 27 Jun 2012 12:21:22 -0400
>From: CCP4 bulletin board <[log in to unmask]> (on behalf of "Bosch, Juergen" <[log in to unmask]>)
>Subject: Re: [ccp4bb] The effect of His-tag location on crystallization
>To: [log in to unmask]
>
> Here's another example:
> http://www.pdb.org/pdb/explore/explore.do?structureId=2F62
> dimer with "His-tag-ears" without His6-tag this
> would not have been possible.
> Jürgen
> On Jun 27, 2012, at 12:04 PM, Brad Bennett wrote:
>
> I think it was an N-terminal RGS-type His tag in
> 3O8Y (human lipoxygenase) that mediated crystal
> contacts with a symmetry related molecule. As I
> recall, this tag composed a B-strand that formed a
> nice interface with a "native" B-strand of the
> symmetry related molecule. Pretty cool...
> -Brad
>
> On Wed, Jun 27, 2012 at 11:00 AM, Phoebe Rice
> <[log in to unmask]> wrote:
>
> With Flp recombinase - DNA complexes, a
> C-terminal His tag triggered a different (but
> sadly not better) crystal form, and the His side
> chains packed against the bases at the end of a
> neighboring DNA duplex.
>
> =====================================
> Phoebe A. Rice
> Dept. of Biochemistry & Molecular Biology
> The University of Chicago
> phone 773 834 1723
> http://bmb.bsd.uchicago.edu/Faculty_and_Research/01_Faculty/01_Faculty_Alphabetically.php?faculty_id=123
> http://www.rsc.org/shop/books/2008/9780854042722.asp
>
> ---- Original message ----
> >Date: Wed, 27 Jun 2012 10:14:58 -0400
> >From: CCP4 bulletin board
> <[log in to unmask]> (on behalf of "R. M.
> Garavito" <[log in to unmask]>)
> >Subject: Re: [ccp4bb] The effect of His-tag
> location on crystallization
> >To: [log in to unmask]
> >
> > Most of the comments you will get will be
> anecdotal
> > in that people will report the successful
> results
> > and do not take the time or effort to
> characterize
> > the less successful results. This often
> occurs
> > because the tagged portion of the protein is
> most
> > often disordered, even in the best crystals.
> Thus,
> > other than saying "tagging on this end
> works, but
> > tagging on that end doesn't," there is
> little more
> > you can say. Each case will be different,
> and it is
> > almost impossible to arrive at any
> generalized
> > conclusion.
> > We prefer C-terminal tagged proteins for a
> number of
> > reasons, but if an N-terminally tagged
> protein
> > crystallizes well, so be it. Of the dozens
> of N-
> > and C-tagged protein structures we have
> solved in my
> > lab and with collaborators, I have only seen
> one
> > case of an ordered His-tag: the His
> residues had
> > coordinated Cd ions, which proved essential
> for
> > getting good crystals. However, beyond that
> there
> > was not much more to say.
> > For your protein and the resulting crystals,
> an
> > N-terminally tagged protein crystallized
> well.
> > Whether you can draw any more conclusions
> from
> > these results depends on characterizing
> crystals of
> > both N- and C-tagged proteins. Just
> assuming that
> > the C-tagged protein is trying to
> crystallize in the
> > same or related crystal form as the N-tagged
> protein
> > is an unwarranted assumption without
> experimental
> > evidence to back it up. That is why most
> groups
> > just run with the winner.
> > Cheers,
> > Michael
> >
> ****************************************************************
> > R. Michael Garavito, Ph.D.
> > Professor of Biochemistry & Molecular
> Biology
> > 603 Wilson Rd., Rm. 513
> > Michigan State University
> > East Lansing, MI 48824-1319
> > Office: (517) 355-9724 Lab: (517)
> 353-9125
> > FAX: (517) 353-9334
> > Email: [log in to unmask]
> >
> ****************************************************************
> > On Jun 26, 2012, at 9:06 PM, weliu wrote:
> >
> > Dear all,
> >
> > We crystallized a protein and found that
> crystal
> > quality greatly depended on the location
> of
> > His-tag. When a His-tag was added at the
> > C-terminus, only crystalline precipitate
> or
> > spherical quasi crystals were grown.
> However, when
> > the His-tag was moved to the N-terminus,
> single
> > crystals were grown under a number of
> conditions,
> > and the best one diffracted to 1.7
> angstrom after
> > optimization. I was wondering if there
> were
> > published reports describing similar
> cases.
> >
> > Thank you in advance
> >
> > Wei Liu
>
> ......................
> Jürgen Bosch
> Johns Hopkins University
> Bloomberg School of Public Health
> Department of Biochemistry & Molecular Biology
> Johns Hopkins Malaria Research Institute
> 615 North Wolfe Street, W8708
> Baltimore, MD 21205
> Office: +1-410-614-4742
> Lab: +1-410-614-4894
> Fax: +1-410-955-2926
> http://web.mac.com/bosch_lab/
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